KR20110132407A - Plant extract compositions for prevention and treatment of influenza - Google Patents

Abstract

A composition affecting at least one of influenza, inflammation, and immune system function, comprising an extract made from Radix isatidis and an extract made from Flos Lonicerae. In an alternative embodiment, the composition further comprises one or more of an extract made from Poria cocos (Schw.) Wolf and an extract made from Herba houttuyniae. A method of treating at least one of influenza, inflammation by administering a therapeutically effective amount of said composition to a patient.

Description

Plant Extract Compositions for Prevention and Treatment of Influenza

Cross reference of related application

This application claims 35 U.S.C. of US Provisional Patent Application No. 61 / 157,049, filed March 3, 2009 entitled "Plant Extract Compositions for the Prevention and Treatment of Influenza." Claims under 119 (e) are hereby incorporated by reference in their entirety.

The present invention relates generally to plant extracts and in particular to the use of plant extracts that affect cold virus infection and modulate the function of the mammalian immune system.

Common cold is a fever virus infection that is acute and typically results in upper respiratory tract inflammation. Common cold is the most common infectious disease affecting humans. Among some other families of viruses that cause a common cold, the most common is rhinoviruses. Estimates indicate that more than one billion cold infections occur in the United States each year. Adults usually catch two to four colds each year, and children typically have colds between six and ten times a year. The annual economic burden resulting from the loss of working days as a result of the cold is estimated to be about $ 5 billion in the United States.

Symptoms of common cold infections include nasal congestion, colds, headaches, teary, burning eyes, fever, and cough. There are a variety of prescription and over-the-counter medications that can be used to treat a common cold. However, these drugs only help relieve the symptoms of a common cold and usually involve a number of side effects such as abdominal pain, nausea, headache, chest pain, diarrhea, constipation, sleepiness and / or dizziness.

Summary of the Invention

Embodiments of the present invention relate to herbal extract compositions for the prevention and treatment of common colds and methods of preparing the same.

Some embodiments of the present invention relate to herbal extract compositions for improving immune system function, particularly when impaired by exogenous stimulation.

Some embodiments of the invention relate to herbal extract compositions for controlling inflammation.

Some embodiments of the invention relate to herbal extract compositions that affect one or more of influenza, inflammation and immune system function.

Some embodiments of the invention relate to a method for the preparation of such herbal extract compositions.

One embodiment of the present invention relates to a herb extract composition comprising a therapeutically effective amount of Radix isatidis, Lonicera japonicum and Poria cocos (Schw.) Wolf.

Another embodiment of the invention is directed to a herbal composition comprising a therapeutically effective amount of panramus and human sinus.

Another embodiment of the present invention is directed to a herbal composition comprising a therapeutically effective amount of rosacea and ghost.

Another embodiment of the invention is directed to a herbal composition comprising a therapeutically effective amount of panramus and Bokyeong.

Another embodiment of the invention is directed to a herbal composition comprising a therapeutic amount of honeysuckle, panlamus muscle and Erba houttuyniae.

Another embodiment of the invention relates to a method of preparing an extract from the aforementioned herbs and combining two or more extracts to provide one of the compositions.

Further embodiments relate to oral dosage forms of the aforementioned herbal compositions including tablets, pills, soft capsules, hard capsules, particulates, powders, concentrates, syrups, liquids, molded balls, and combinations thereof.

Another embodiment relates to a method of treating one or more of influenza, inflammation and immune system function by administering a therapeutically effective amount of the aforementioned herbal composition.

Embodiments of the present invention comprise a composition comprising a mixture of herbal extracts selected for the prevention and treatment of influenza, inflammation as well as for affecting and / or regulating immune system function, methods of making such compositions, and the use of the compositions It is about a method for.

In one embodiment, the present invention relates to a composition comprising an extract from Panlamus muscle and Bokryeong. In a preferred embodiment, the panlammus and Bokryeong extract are present in a ratio of about 1.0: 0.2 to about 1.0: 3.0. Alternatively, the composition may further comprise Echochocho extract.

In another embodiment, the present invention is directed to a composition comprising extracts from phosphorus and bokyeong. In a preferred embodiment, the phosphorus and Bokryeong extracts are present in a ratio of about 1.0: 0.2 to about 1.0: 3.0. Alternatively, the composition may further comprise Echochocho extract.

In another embodiment, the present invention is directed to a composition comprising individual extracts from panramus and phosphorus. In a preferred embodiment, the panlammus and phosphorus extract are present in a ratio of about 1.0: 0.2 to about 1.0: 3.0. Alternatively, the composition may further comprise Echochocho extract.

A further embodiment relates to a composition comprising an extract made from panlamus muscle, an extract made from human sinus, and an extract made from Bokryeong. In a preferred embodiment, the panlam root, honeysuckle and Bokyeong extract are present in a ratio of about 1.0: 1.0: 2.1.

Another embodiment of the invention is directed to a composition comprising extracts made individually from Panramus, Phosphorus, Bokryeong, and Eochocho.

Panlam Geun is a traditional Chinese herbal medicine derived from the roots of Daejeon (Isatis tinctoria L) and Songram (Isatis Indigotica Fort), commonly known as Daechung. For the purposes described herein, it is appropriate to use dry roots, leaves, and stems of marlin plants to prepare panramus extract. In one embodiment, the extract may be derived from about 5 to 20 g of panramus dry root. Alternatively, the extract can be prepared from fresh or dried flowers, leaves, stems, and seeds.

Extracts of Flos Lonicerae, also referred to as Lonicera japonicum, Japanese Honeysuckle, White Honeysuckle, or Chinese Honeysuckle, may be appropriately prepared from fresh or dried flowers, leaves, and stems. In one embodiment, the extract may be derived from about 5-20 g of dried flowers.

Fuling is a land tree rot fungus. Its subterranean fungus is commonly used in oriental medicine. Bokryeong extract can be prepared from dry bacterium. In one embodiment, the extract may be derived from about 5-20 g of dry bacterium. Alternatively, the extract can be prepared from fresh fungus.

Eochocho is an herbal medicine produced from a part of the ground of Houttuynia cordata Thunb., For example, shoots, leaves and stems. Echochocho extract can be prepared from some of the fresh or dried plants. In one embodiment, the extract may be derived from about 10-30 g of dry plant parts.

Those skilled in the art will appreciate that the extract may be derived from other amounts of the aforementioned herbs. In addition, those skilled in the art will recognize that these herbal extracts contain multiple compounds, all of which are used in the preparation of the aforementioned compositions.

Methods for preparing each extract are generally:

(a) immersing the selected plant parts and / or components with an appropriate volume of aqueous solvent at ambient room temperature for a period of time selected from about 15 minutes to about 24 hours to produce an extraction mixture;

(b) filtering the extraction mixture to produce an aqueous filtrate;

(c) obtaining an aqueous filtrate; And

(d) concentrating, i.e., dehydrating, the aqueous filtrate to a concentrated liquid or syrup-like density.

In another embodiment, the method further comprises (e) dehydrating the aqueous filtrate to about semisolid or pool-like density. In a further alternative embodiment, the method further comprises (f) dehydrating the aqueous filtrate to dry density, for example particulates and powder.

Suitable aqueous solvents for use in the process are water. Alternative aqueous solvents may include organic solvents such as: ethanol, methanol, isopropyl alcohol, ethyl acetate, ethyl ether, acetonitrile, methylene chloride, hexane, acetic acid, and combinations thereof. In alternative embodiments, the aqueous solvent may further comprise an inorganic acid or inorganic base alone or in combination with one or more selected organic solvents. In a further alternative embodiment, the aqueous solvent is mixed with the plant and / or plant components for a period of time to soak.

In another embodiment, the extract solvent is a mixture of organic solvents.

In an alternative embodiment, the method may further comprise heating to a range from about higher than about ambient room temperature to about 110 ° C. during the soaking period of step (a). In a further alternative embodiment, when not heated during the soaking period of step (a), the process comprises aqueous solvents and plants for a period of time ranging from about 15 minutes to about 2 hours after the soaking period to produce an extract mixture. The method may further include heating the mixture.

In alternative embodiments, extraction may also be performed using methods known in the art, including traditional decoction. Alternatively, the extract can be further purified using a suitable chromatography column such as silica gel and Sephadex LH-20. In further alternative embodiments, the plant material may be extracted using a supercritical carbon dioxide device and process, and / or a reflux extraction process.

In one embodiment, the extract is formulated in an oral dosage form. The extract may be encapsulated in soft capsules or hard capsules. In alternative embodiments, the extract may be formulated as a concentrate, syrup and liquid. In another alternative embodiment, the extract is mixed with one or more pharmaceutically acceptable carriers and / or excipients to produce the resulting mixture suitable for forming into tablets, pills, soft capsules, hard capsules, and combinations thereof.

In another alternative embodiment, the extract can be formulated as a dry powder. In another alternative embodiment, the extract can be formulated as microparticles. In an alternative embodiment, the extract can be formulated as a cast ring with a flexible clay-like density according to traditional herbal medicine. Such cast rings are easily bent, easily made, and made into rings of various sizes depending on the patient's ability to consume larger or smaller amounts of the cast ring.

The compositions of the present invention and methods for their preparation and use are described in more detail in the Examples below.

Example

Preparation of Extracts:

Extracts for panramus and sterling were separately prepared using the previously described method and 70% ethanol extraction in water. The Fuling herb was extracted by dissolution in 0.6M NaOH solution, neutralization by addition of acetic acid, and then precipitation through addition of ethanol. Each extract was dried in powder form. The extract powders were combined in mass to mass ratio as shown in Table 1 and labeled with "AB" and "ABC", respectively:

Extract powder AB ABC A (gold and silver) One One B (Pectus Root) One One C (Restoration) 0 2.12

material:

Virus Seeds: Type 1 FM 1 influenza virus adapted to mouse lungs was obtained from the Detection Department of Chinese Medicine and Biological Products. The toxicity of the FM1 virus was enhanced in mice and then passed twice through Embryo urinary cysts. LD50 was 5.2.

Virus Dilution: The virus seed culture was dissolved in water and diluted with sterile saline to 0.05 ml per 15 LD50 before starting the experiment.

Mouse Infection: Mice were infected by dropping influenza virus intranasally with 4 drops, about 0.05 ml, for each mouse.

Dosing: Oral dosing began 6 hours after viral administration was performed. The oral dosage dose for each animal was calculated on a 20 ml / kg basis.

Example  One: FM1  Inhibition of Influenza Viruses

Effective inhibition of FM1 influenza virus was determined by measuring lung index values for a series of treatment groups.

The low lung index calculated for the virus-infected animal control indicates that their lungs were more affected than the lungs of the normal control animal group. A probability below 0.05 indicates that the herbal composition or drug was effective in treating the virus. The results are shown in Table 2 below.

Between 15 and 15 grams of NIH mice, in grades without specific pathogens, were used in the experiments described below. Half of the test animals in each test group were male. Breeding concentration was 23 ± 2 ℃.

Mice were randomly divided into 9 groups with at least 10 mice in each group: (1) normal control, (2) virus-infected control, (3) AB high dose (15 g / kg), ( 4) AB intermediate dose (7.5 g / kg), (5) AB low dose (3.75 g / kg), (6) ABC high dose (13.0 g / kg), (7) ABC intermediate dose (7.66) g / kg), (8) ABC low dose (3.83 g / kg), and (9) ribavirin group (0.07 g / kg).

Treatment of the placebo and extract compositions is administered intragastrically once a day for a total of three days. Normal control mice and virus-infected control mice received the same volume of distilled water.

Normal control groups included mice not infected with virus. The normal control group was treated via placebo, distilled water administration. Virus-infected control groups include mice infected with FM1 virus. The virus-infected control group was treated through the administration of placebo, distilled water. The ribavirin group provides a baseline to explain the effective level of treatment of type 1 FM1 of influenza virus.

Determination of Lung Index: Mice were sacrificed 4 days after viral infection treatment. Drinking water was removed and fasted for at least 4 hours before the sacrifice of the test animals. The mice were then weighed and the lungs were removed after cervical dislocation. Lung damage levels were recorded. The lungs were then washed twice with 0.9% standard saline, dewatered and reweighed by blotting paper. An increase in lung index values indicates an increase in the degree of lung disease.

Lung index and inhibition rate of lung index were calculated by the following formula:

Lung index = (mouse lung weight / mouse weight) * 10

Inhibition rate of lung index = ((average lung index of control group-average lung index of experimental group) / average lung index of control group) × 100

Mouse Lung Index Value (x ± se) group Dose
(g / kg) animal
(n) weight
(g) Lung index
(g / g) Inhibition rate of lung index
(%) P-value
(Dunnet t both sides black) Normal control group - 11 17.04 ± 1.00 ^** 0.94 ± 0.11 ^** - 0.000 ^** Virus-infected control group - 10 11.86 ± 0.84 1.76 ± 0.15 - - Ribavirin 0.07 11 14.94 ± 1.64 ^** 1.19 ± 0.30 ^** 32.38 0.000 ^** AB 15.0 12 12.85 ± 0.56 1.41 ± 0.12 ^** 19.88 0.005 ^** 7.5 11 12.60 ± 1.24 1.34 ± 0.26 ^** 25.56 0.001 ^** 3.75 11 12.82 ± 1.35 1.46 ± 0.22 ^* 17.04 0.034 ^* ABC 13.0 11 12.81 ± 1.81 1.41 ± 0.28 ^** 19.88 0.008 ^** 7.66 10 12.24 ± 0.57 1.40 ± 0.29 ^** 20.45 0.007 ^** 3.83 11 11.99 ± 0.58 1.53 ± 0.30 13.06 0.161

Note: compared with virus control group ^* : P <0.05; ^** : P <0.01

The results shown in Table 2 above show that the mouse weight of the virus-infected group was significantly reduced and the lung index was significantly increased compared to the normal control group, indicating that the virus infected animal model was well established. In addition, the low lung index for the virus-infected control indicates that their lungs were more affected than the lungs of the normal control animal group. A probability below 0.05 indicates that treatment of the herbal composition or baseline drug was effective.

Mice in the groups receiving high AB, medium AB and low AB extract doses and high ABC, and medium ABC extract doses reduced lung disease as indicated by a decrease in lung index in comparison with the virus-infected control group. Showed. Furthermore, when comparing the mice in the high AB, intermediate AB, high ABC and intermediate ABC extract dose groups with the group of ribavirin treated mice, each AB extract dose and ABC extract dose were similar to those treated with ribavirin. Obviously, it is clear that the influenza virus is suppressed, as is apparent with similar inhibition of the lung index. The data in Table 2 clearly demonstrate that AB and ABC extracts can significantly inhibit type 1 FM1 of influenza virus.

Example  2: Analysis of Anti-inflammatory Effects of Extract Compositions

The anti-inflammatory effects of the extract compositions AB and ABC were determined by measuring the permeability of intraperitoneal capillaries for a series of treatment groups.

NIH mice weighing 20 ± 2 g were used for this experiment. Each test group had the same number of male and female mice. The breeding temperature was 23 ± 2 ° C. Mice were randomly divided into 9 groups with 10 mice in each group: (1) blank control, (2) model control, (3) AB high dose (15 g / kg), (4) AB Medium dose (7.5 g / kg), (5) AB low dose (3.75 g / kg), (6) ABC high dose (13.0 g / kg), (7) ABC intermediate dose (7.66 g / kg) ), (8) ABC low dose (3.83 g / kg), and (9) sodium salicylate group (0.15 g / kg).

The blank control group included mice that were not infected with virus, treated with placebo. Model control groups include mice infected with FM1 virus, treated with placebo. The sodium salicylate group provides a baseline to explain the effective level of treatment of type 1 FM1 of influenza virus.

Acetic standard saline solution was prepared by dissolving 0.4% glacial acetic acid in 50 ml of standard saline.

A 2% Evans Blue standard saline solution was prepared by dissolving 2 g of Evans Blue in 100 ml standard saline.

Treatment of placebo and AB and ABC extract compositions was administered once daily to the stomach for a period of 3 days. The blank control group was treated through the administration of placebo, saline solution. Model control groups were treated by administration of placebo, saline solution.

On the morning of the fourth day, one hour after the last treatment dose, each mouse in each group was injected with 2% Evans Blue standard saline via the tail vein (0.1 mL / lO g). The blank group was then injected 0.2 ml of standard saline into the abdominal cavity. The rest of the group received 0.2 ml of 0.8% acetic acid standard saline intraperitoneally.

About 20 minutes after the saline injection, each mouse was sacrificed by cervical distal bone. The abdominal skin and muscles were then excised. Thereafter, peritoneal macrophages were collected by washing the abdominal cavity with 5 ml of standard saline. The resulting wash solution was then collected and centrifuged at 3000 rpm for about 15 minutes. The supernatant was then collected and the absorbance or optical density (OD) of the supernatant was measured at 570 nm using a spectrophotometer. Experimental data were analyzed. Data were evaluated using the t-test to determine statistically significant differences between the results of each group. The anti-inflammatory effects of the AB and ABC herbal extract compositions and sodium salicylate were considered effective when P was less than 0.05. The results are shown in Table 3.

Optical density is an indicator of the level of permeability of intraperitoneal capillaries. As the intraperitoneal macrophage level increases, the optical density increases, indicating an increase in permeability in the intraperitoneal capillaries. Administration of acetic acid increases permeability of intraperitoneal capillaries.

Anti-inflammatory effect group Dose (g / kg) animal OD value of hemolysin in serum Blank Control - 10 0.299 ± 0.086 Model control group - 10 0.341 ± 0.023 ^ΔΔ Sodium salicylate 0.15 10 0.305 ± 0.060 ^** AB 3.75 10 0.323 ± 0.06 ^* AB 7.5 10 0.316 ± 0.067 ^* AB 15 10 0.295 ± 0.086 ^** ABC 3.83 10 0.312 ± 0.018 ^** ABC 7.66 10 0.325 ± 0.058 ^* ABC 13 10 0.310 ± 0.043 ^**

^{Compared with ΔΔ} blank group, P < ^O.01

^* Comparison with model group: P <O.05

^** Comparison with model group: P <0.01

The results shown in Table 3 clearly show that the optical density of the model group is significantly higher than that of the blank group. This indicates that the animal model is well established.

In comparison with the model control group, the reduced optical density of the groups treated with the AB and ABC herbal compositions indicates the ability of these herbal compositions to reduce the permeability of intraperitoneal capillaries, and thus the respective of the AB and ABC herbal composition groups Dose level anti-inflammatory effects have been demonstrated.

When mice in the high AB, low ABC and high ABC extract dose groups were compared with mice in the sodium salicylate treated group, each of the aforementioned ABC extract doses and AB extract doses of sodium salicylate was compared. It is evident that an anti-inflammatory effect similar to the treatment was demonstrated.

The data in Table 3 clearly show that the AB and ABC extracts had significant anti-inflammatory effects and their efficacy was similar to sodium salicylate.

Example  3: Carrageenan  Induced by Rat  Analysis of the anti-inflammatory effect of the extract composition in paw swelling

The anti-inflammatory effects of the extract compositions AB and ABC were determined by measuring the swelling of the rat paw after administration of carrageenan for a series of treatment groups.

Rats weighing 200 ± 20 g were used in this experiment. Each test group included the same number of male and female animals. The breeding temperature was 23 ± 2 ° C. The rats were divided into eight groups of 10 rats each: (1) model control, (2) AB high dose (15 g / kg), (3) AB intermediate dose (7.5 g / kg), ( 4) AB low dose (3.75 g / kg), (5) ABC high dose (13.0 g / kg), (6) ABC intermediate dose (7.66 g / kg), (7) ABC low dose (3.83 g / kg), (8) sodium salicylate group (0.15 g / kg), and (9) blank control. In this experiment, there was no blank control because there was no swelling of the animal's paw when no carrageenan was administered.

The method for performing this experiment followed the guidelines outlined in the Manual on Studies of New Medicines of TCM by the Drug Administration Ministry of Health P.R.C. The method outlined in the manual details the following steps:

1. With a ballpoint pen, draw a clear horizontal line located on the upper ankle of the left hind paw of each rat;

2. Measure the paw volume of each rat;

3. Except for those in the blank control group, each rat is injected subcutaneously with a dose of carrageenan (0.05 ml / foot) in the ulna of the hind legs;

4. Once per day for a period of four days, orally administer either the AB extract, the ABC extract, or the treatment of sodium salicylate, according to the aforementioned dosages. Rats in the model control group and the blank were injected with the same volume of saline;

5. On the morning of the fourth day, 1 hour after the last treatment, carrageenan (0.05 mL / foot) is injected subcutaneously into the ulna of each hind leg;

6. Measure foot volume 1, 2, 3 and 4 hours after the second carrageenan administration, respectively; And

7. Calculate the paw book percentage for each rat.

Bookkeeping percentage was calculated by the following formula:

Swelling percentage = ((swelling volume of rat foot after inflammation-swelling volume of rat foot before inflammation) / swelling volume of rat foot before inflammation) × 100%

Experimental data were analyzed using the t-test to determine statistically significant differences between the results of each group. There was no swelling in the paws of the rats of the blank control group. The anti-inflammatory effects of the AB and ABC herbal extract compositions and sodium salicylate were considered effective when P was less than 0.05. The results are shown in Table 4 below.

Compared with the model group, the low and medium AB extract doses demonstrated a significant anti-inflammatory effect at 4 hours, while the high ABC extract doses were at 1, 2, 3 and 4 hours (p <0.01) time points, respectively. It showed a significant anti-inflammatory effect. The data in Table 4 clearly show that the low AB, middle AB and high ABC extracts have significant anti-inflammatory effects and their efficacy is similar to sodium salicylate.

Anti-inflammatory Effects in Carogienin-induced Foot Boogie group Dose
(g / kg) animal
(n) Foot swelling (ml) 1h 2h 3h 4h Model control group - 10 0.865 ± 0.208 0.820 ± 0.174 0.720 ± 0.199 0.605 ± 0.215 Sodium salicylate 0.15 10 0.490 ± 0.202 ^** 0.405 ± 0.195 ^** 0.265 ± 0.153 ^** 0.170 ± 0.136 ^** AB 15 10 0.752 ± 0.168 0.745 ± 0.182 0.702 ± 0.146 0.612 ± 0.113 7.5 10 0.700 ± 0.175 0.705 ± 0.144 0.615 ± 0.181 0.455 ± 0.101 ^** 3.75 10 0.710 ± 0.133 0.690 ± 0.149 0.655 ± 0.185 0.420 ± 0.178 ^** ABC 13 10 0.575 ± 0.106 ^** 0.555 ± 0.119 ^** 0.540 ± 0.158 ^* 0.375 ± 0.130 ^** 7.66 10 0.615 ± 0.113 ^* 0.596 ± 0.108 ^* 0.563 ± 0.115 ^* 0.512 ± 0.132 ^* 3.83 10 0.685 ± 0.106 ^* 0.636 ± 0.118 ^* 0.589 ± 0.162 ^* 0.554 ± 0.123 ^* Blank Control - 10 - - - -

^* P <0.01 compared with model group

^** Compared to model group p <0.05

Example  4. Analysis of anti-inflammatory effects of extract compositions in immune function

The anti-inflammatory effects of the extract compositions AB and ABC were determined through antibody level tests circulating in both red blood cells for a series of treatment groups.

Mice weighing 20-25 g were used in this study. Each group included the same number of male and female animals. The breeding temperature was 23 ± 2 ° C. Mice were divided into nine groups, each containing 10 rats: (1) blank control, (2) model control, (3) AB high dose (15 g / kg), (4) AB intermediate dose ( 7.5 g / kg), (5) AB low dose (3.75 g / kg), (6) ABC high dose (13.0 g / kg), (7) ABC intermediate dose (7.66 g / kg), (8 ) ABC low dose (3.83 g / kg), and (9) levamisol (0.03 g / kg).

Treatment of the placebo and extract compositions was administered to each mouse by gastric infusion for a period of 7 days. On days 2, 4, and 6, each mouse was intraperitoneally injected with 50 mg / 10 ml / kg of cyclophosphamide, except for those of the blank control group. Mice in the blank control group were injected with the same volume of saline intraperitoneally. Three days after administration of the last treatment (day 7), all mice were immunized by intraperitoneal injection of 0.2 ml of both red blood cell suspensions (5% of both red blood cells in saline) respectively. Five days after immunization, the mice's eyes were removed under anesthesia. Blood was collected, then serum was separated and the serum diluted approximately 100-fold with saline. Then 1 ml of diluted serum was mixed with 0.5 ml of 5% sheep erythrocyte suspension and 0.5 ml of 10% complement. Complement was made by mixing serum from three rats with saline solution at a ratio of 1:10. The resulting serum solution was stored in an incubator at a temperature of about 37 ° C. for a period of about 30 minutes. The resulting serum solution was then placed in the refrigerator at a temperature of about 0 ° C. to stop further reactions. The serum solution was then centrifuged to collect the supernatant. Color comparison of the supernatants was performed at 540 nm using spectrophotometer UV-721. Absorbance for each supernatant sample was recorded as an index of serum hemolysis level. The results are shown in Table 5 below.

The results indicate that the level of hemolysine serum specific antibodies in the model group was significantly reduced by cyclophosphamide compared to the blank group (P <0.01), indicating that the animal model was well established.

Optical density is an indicator of the antibody level of hemolysin. In comparison to the blank group, the model group that received cyclophosphamide had a significantly low OD value, indicating that the antibody level of hemolysin was significantly reduced by cyclophosphamide. Administration of the herbal extract composition treatment as described above has been shown to significantly increase the levels of antibodies of hemolysin in immunodeficient mice induced by cyclophosphamide.

In comparison to the model group, the AB and ABC extract compositions significantly improved immune function in immunodeficient mice induced by cyclophosphamide and increased phagocytic capacity of the reticuloendothelial system. High AB doses and high, medium and low ABC doses had similar efficacy to levamisol in improving immune function in mice.

Effect of Sheep Red Blood Cells on Circulating Antibody Level Testing group Dose
g / kg Number of animals OD value of hemolysin Blank Control - 10 0.74 + 0.009 Model control group - 10 0.56 ± 0.004 ^ΔΔ Levamisol 0.03 10 0.75 ± 0.006 ^** AB 3.75 10 0.68 ± 0.006 ^* AB 7.5 10 0.66 ± 0.008 ^* AB 15 10 0.72 ± 0.006 ^** ABC 3.83 10 0.70 ± 0.018 ^** ABC 7.66 10 0.69 ± 0.008 ^** ABC 13 10 0.73 ± 0.043 ^**

^Compared with ^ΔΔ blank, P < ^O.01

^** Compared to model, P <O.01

^* Compared to model, P <0.05

Example  6: peritoneum in mice Macrophage In phagocytosis  Analysis of the Effect of Extract Compositions

Anti-inflammatory effects of extract compositions AB and ABC were determined through analysis of peritoneal macrophage phagocytosis in mice for a series of treatment groups.

Mice weighing 20-25 g were used in this study. Each group included the same number of male and female animals. Mice were randomly divided into 9 groups of 10 mice each: (1) blank control, (2) model control, (3) AB high dose (15 g / kg), (4) AB intermediate dose (7.5 g / kg), (5) AB low dose (3.75 g / kg), (6) ABC high dose (13.0 g / kg), (7) ABC intermediate dose (7.66 g / kg), ( 8) ABC low dose (3.83 g / kg), and (9) levamisol hydrochloride group. The blank and model control groups were administered orally with the same volume of saline.

Treatment was administered to mice by gastric infusion for 7 days. On days 2, 4, and 6, each mouse was intraperitoneally injected with 50 mg / 10 ml / kg of cyclophosphamide, except for those of the blank control group. The blank group received an equal volume of saline solution intraperitoneally. Mice were injected with Indian ink through the tail vein at a dose of 0.1 mL / 1O g. Venous blood was collected from the orbit at 2 and 10 minutes after injection, respectively. 20 μl of venous blood were mixed thoroughly with 2 ml of 0.1% sodium carbonate solution. The optical density (OD) for each sample was then measured at 600 nm by spectrophotometer UV-752.

Phagocyte count K and phagocytic count index α were calculated using the following formula:

Phagocyte count K = (logOD ₁ -logOD ₂ ) / (t ₂ -t ₁ )

Phagocyte counting index α = W / WLS

t: time, W: mouse weight, WLS: weight of liver and spleen.

Table 6 below shows the phagocytic count (K) and phagocytic count index (α) for each group. The phagocytic count (K) and phagocytic count index (α) for the model group were significantly reduced compared to the blank control group, indicating that the animal model was well established. Each of the AB and ABC herb compositions had significant antagonism in the inhibition of cellular immunity induced by cyclophosphamide, and phagocytic count (K) and phagocyte count index near normal levels as indicated by the blank control group level. (α) was recovered. All AB and ABC herbal compositions demonstrated improved immune function in cyclophosphamide induced immunodeficiency mice and their efficacy was similar to that of the levamisol group.

Comparison with blank group: ^□□ P <0.01

Compared with model, ^** P <0.01,

Comparison with the model ^* P <0.05

While preferred embodiments of the invention have been shown and described herein, those skilled in the art will clearly recognize that such embodiments are provided by way of illustration only. Numerous variations, changes, and substitutions will occur to those skilled in the art without departing from the invention. It should be understood that various alternatives to the embodiments of the invention described herein may be used in the practice of the invention. The following claims define the scope of the present invention, and the methods and configurations within the scope of these claims, and their equivalents, are intended to be covered thereby.

Claims (21)

A composition affecting at least one of influenza, inflammation, and immune system function, comprising an extract made from Radix isatidis and an extract made from Flos Lonicerae. The composition of claim 1, wherein the composition further comprises an extract prepared from Poria cocos (Schw.) Wolf. The composition of claim 2, wherein the composition further comprises an extract prepared from Herba houttuyniae. A composition affecting one or more of influenza, inflammatory and immune system functions, including extracts prepared from panlamus muscle and extracts prepared from Bokryeong. The composition of claim 4, wherein the composition further comprises an extract prepared from fishery herb. A composition affecting at least one of influenza, inflammation, and immune system function, comprising an extract made from gold and silver and an extract made from Bokyeong. 7. The composition of claim 6, wherein the composition further comprises an extract prepared from fishery herb. The composition of claim 1, wherein the composition further comprises a pharmaceutically acceptable carrier and / or excipient. The composition of claim 1, wherein the composition is in oral dosage form. 10. The composition of claim 9, wherein the oral dosage form is selected from the group comprising tablets, pills, soft capsules, hard capsules, particulates, powders, concentrates, syrups, liquids, cast rings and combinations thereof. Immersing the plant components in an aqueous solvent at a temperature selected from the range of about 20 ° C. to about 110 ° C., thereby preparing an herbal extract mixture;
Separating the liquid fraction from the herbal extraction mixture and filtering the liquid fraction to produce an aqueous herbal filtrate;
Dewatering the aqueous herbal filtrate to a syrup-like density; And
Preparing a composition comprising the dehydrated herbal filtrate and an acceptable carrier or excipient thereof,
A method of making a composition that affects one or more of influenza, inflammation, and immune system function. 12. The composition of claim 11, wherein the composition is formulated as an oral dosage form selected from the group comprising tablets, pills, soft capsules, microparticles, powders, concentrates, syrups, liquids, cast pills and combinations thereof. Way. 12. The method of claim 11, wherein said plant component is selected from the group consisting of panlamus, gilt, bokyeong, and estuary. The method of claim 11, wherein the aqueous solvent is at least one organic solvent selected from the group comprising ethanol, methanol, isopropyl alcohol, ethyl acetate, ethyl ether, acetonitrile, methylene chloride, hexane, acetic acid, and combinations thereof. And a mixture of water. The method of claim 11, wherein the aqueous solvent further comprises a mixture of water with at least one of an inorganic acid, an inorganic base, and combinations thereof. 12. The method of claim 11, further comprising dewatering and processing the aqueous herbal filtrate to produce a dried powder. A method of treating at least one of influenza, inflammation by administering to a patient a therapeutically effective amount of the composition of claim 1. The composition of claim 1, wherein the panlam root extract and the sterling extract are present in a ratio of about 1.0: 0.2 to about 1.0: 3.0. 5. The composition of claim 4, wherein the Panlam root extract and the Bokryeong extract are present in a ratio of about 1.0: 0.2 to about 1.0: 3.0. The composition of claim 6, wherein the sterling silver extract and the bokyong extract are present in a ratio of about 1.0: 0.2 to about 1.0: 3.0. 3. The composition of claim 2, wherein the panlam root extract, the sterling silver extract, and the bokyong extract are present in a ratio of about 1.0: 1.0: 2.1.